Please use this identifier to cite or link to this item: https://doi.org/10.1186/s13287-017-0538-x
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dc.titleCritical attributes of human early mesenchymal stromal cell-laden microcarrier constructs for improved chondrogenic differentiation
dc.contributor.authorLin, Y.M
dc.contributor.authorLee, J
dc.contributor.authorLim, J.F.Y
dc.contributor.authorChoolani, M
dc.contributor.authorChan, J.K.Y
dc.contributor.authorReuveny, S
dc.contributor.authorOh, S.K.W
dc.date.accessioned2020-10-23T04:47:23Z
dc.date.available2020-10-23T04:47:23Z
dc.date.issued2017
dc.identifier.citationLin, Y.M, Lee, J, Lim, J.F.Y, Choolani, M, Chan, J.K.Y, Reuveny, S, Oh, S.K.W (2017). Critical attributes of human early mesenchymal stromal cell-laden microcarrier constructs for improved chondrogenic differentiation. Stem Cell Research and Therapy 8 (1) : 93. ScholarBank@NUS Repository. https://doi.org/10.1186/s13287-017-0538-x
dc.identifier.issn17576512
dc.identifier.urihttps://scholarbank.nus.edu.sg/handle/10635/179501
dc.description.abstractBackground: Microcarrier cultures which are useful for producing large cell numbers can act as scaffolds to create stem cell-laden microcarrier constructs for cartilage tissue engineering. However, the critical attributes required to achieve efficient chondrogenic differentiation for such constructs are unknown. Therefore, this study aims to elucidate these parameters and determine whether cell attachment to microcarriers throughout differentiation improves chondrogenic outcomes across multiple microcarrier types. Methods: A screen was performed to evaluate whether 1) cell confluency, 2) cell numbers, 3) cell density, 4) centrifugation, or 5) agitation are crucial in driving effective chondrogenic differentiation of human early mesenchymal stromal cell (heMSC)-laden Cytodex 1 microcarrier (heMSC-Cytodex 1) constructs. Results: Firstly, we found that seeding 10 × 103 cells at 70% cell confluency with 300 microcarriers per construct resulted in substantial increase in cell growth (76.8-fold increase in DNA) and chondrogenic protein generation (78.3- and 686-fold increase in GAG and Collagen II, respectively). Reducing cell density by adding empty microcarriers at seeding and indirectly compacting constructs by applying centrifugation at seeding or agitation throughout differentiation caused reduced cell growth and chondrogenic differentiation. Secondly, we showed that cell attachment to microcarriers throughout differentiation improves cell growth and chondrogenic outcomes since critically defined heMSC-Cytodex 1 constructs developed larger diameters (2.6-fold), and produced more DNA (13.8-fold), GAG (11.0-fold), and Collagen II (6.6-fold) than their equivalent cell-only counterparts. Thirdly, heMSC-Cytodex 1/3 constructs generated with cell-laden microcarriers from 1-day attachment in shake flask cultures were more efficient than those from 5-day expansion in spinner cultures in promoting cell growth and chondrogenic output per construct and per cell. Lastly, we demonstrate that these critically defined parameters can be applied across multiple microcarrier types, such as Cytodex 3, SphereCol and Cultispher-S, achieving similar trends in enhancing cell growth and chondrogenic differentiation. Conclusions: This is the first study that has identified a set of critical attributes that enables efficient chondrogenic differentiation of heMSC-microcarrier constructs across multiple microcarrier types. It is also the first to demonstrate that cell attachment to microcarriers throughout differentiation improves cell growth and chondrogenic outcomes across different microcarrier types, including biodegradable gelatin-based microcarriers, making heMSC-microcarrier constructs applicable for use in allogeneic cartilage cell therapy. © 2017 The Author(s).
dc.rightsAttribution 4.0 International
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.sourceUnpaywall 20201031
dc.subjectcollagen type 2
dc.subjectDNA
dc.subjectGag protein
dc.subjectCytodex
dc.subjectdextran
dc.subjectmicrosphere
dc.subjectArticle
dc.subjectcell adhesion
dc.subjectcell confluency
dc.subjectcell count
dc.subjectcell density
dc.subjectcell differentiation
dc.subjectcell expansion
dc.subjectcell growth
dc.subjectcell size
dc.subjectcellular parameters
dc.subjectcentrifugation
dc.subjectchondrogenesis
dc.subjectcontrolled study
dc.subjecthuman
dc.subjecthuman cell
dc.subjectmesenchymal stroma cell
dc.subjectmicrocarrier culture
dc.subjectprotein expression
dc.subjectprotein synthesis
dc.subjectsurface area
dc.subjectadverse device effect
dc.subjectcell culture
dc.subjectchemistry
dc.subjectcytology
dc.subjectdrug effects
dc.subjectmesenchymal stroma cell
dc.subjectprocedures
dc.subjecttissue engineering
dc.subjecttissue scaffold
dc.subjectCell Differentiation
dc.subjectCells, Cultured
dc.subjectChondrogenesis
dc.subjectDextrans
dc.subjectHumans
dc.subjectMesenchymal Stromal Cells
dc.subjectMicrospheres
dc.subjectTissue Engineering
dc.subjectTissue Scaffolds
dc.typeArticle
dc.contributor.departmentOBSTETRICS & GYNAECOLOGY
dc.contributor.departmentDUKE-NUS MEDICAL SCHOOL
dc.description.doi10.1186/s13287-017-0538-x
dc.description.sourcetitleStem Cell Research and Therapy
dc.description.volume8
dc.description.issue1
dc.description.page93
dc.published.statePublished
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